A structure-function analysis of P2 integrase
(English)Manuscript (preprint) (Other academic)
Bacteriophage P2 integrase catalyzes site-specific recombination between the phage DNA and the host chromosome thereby promoting integration or excision of the phage genome. P2 integrase belongs to the large tyrosine family of integrases that shows little sequence identity besides some conserved boxes and patches in the catalytic domain. However, the overall structure of the tyrosine family of integrases seems to be similar. Phage integrases have the potential as tools for site-specific gene insertions into eukaryotic genomes provided that target sequences are available. To elucidate the possibility of evolving the P2 integrase to accept new targets, we have in this work initiated a structure-function analysis of the P2 integrase using two approaches based on a comparison of the predicted secondary structure of P2 integrase with that determined for the lambda integrase. First, we have made hybrids between P2 integrase and the related WΦ integrase that has a different host DNA target, to locate the region promoting specificity between the integrases. This, however, has not been possible, the N-terminal domains can be exchanged without losing biological activity and this will not affect the specificity. All other hybrids made were biological inactive. Next we have made an alanine scanning of the alpha helices believed to be involved in specific interactions with the target, and four amino acids have been identified as candidates for sequence-specific interactions with the core.
site-specific recombination, integrase, tyrosine recombinase, bacteriophage
Genetics Biochemistry and Molecular Biology
Research subject Molecular Genetics
IdentifiersURN: urn:nbn:se:su:diva-38927OAI: oai:DiVA.org:su-38927DiVA: diva2:317571
Authors 1 and 2: Equal contribution to this work2010-05-042010-05-042010-11-17Bibliographically approved